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Application of pnliprp3 gene and its expression product in the diagnosis and treatment of squamous cell carcinoma of the tongue

A technology of gene expression and tongue squamous cell carcinoma, applied in the field of tumor diagnosis and prognosis prediction, can solve the problems of wide surgical resection, affecting the quality of life of patients, and large lesion area.

Active Publication Date: 2020-03-31
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the size of the lesion is large and the scope of surgical resection is wide, which will seriously affect the quality of life of patients after surgery.

Method used

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  • Application of pnliprp3 gene and its expression product in the diagnosis and treatment of squamous cell carcinoma of the tongue
  • Application of pnliprp3 gene and its expression product in the diagnosis and treatment of squamous cell carcinoma of the tongue
  • Application of pnliprp3 gene and its expression product in the diagnosis and treatment of squamous cell carcinoma of the tongue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening of differentially expressed genes

[0063] 1. Experimental materials

[0064] Tissue specimens from 5 cases of tongue squamous cell carcinoma were collected from oral and maxillofacial surgery patients, including 2 cases of well-differentiated squamous cell carcinoma, 2 cases of moderately differentiated squamous cell carcinoma, and 1 case of poorly differentiated squamous cell carcinoma, including 2 males and 3 females. At the same time, the normal tissue around the tumor of each case > 5 cm was selected as the self-control. All patients had not undergone chemotherapy, radiotherapy, biological therapy and other treatments for tumors before treatment. A portion of the tissue was immediately placed in liquid nitrogen for storage.

[0065] 2. RNA extraction, cDNA synthesis

[0066] Total RNA was extracted with Trizol RNA reagent (Invitrogen Company), and total RNA was identified by UV spectrophotometer (ND-1000, NanoDrop Company) and agarose gel ele...

Embodiment 2

[0075] Example 2 Validation of differentially expressed genes in large samples

[0076] Based on the results of the primary screening on the chip, we selected the PNLIPRP3 gene for large-scale verification.

[0077] 1. Sample collection

[0078] According to the method of Example 1, 45 cases of tongue squamous cell carcinoma tissues and 45 normal tissues were collected.

[0079] 2. Validation at the mRNA level

[0080] 2.1 Extract tissue RNA

[0081] The steps are the same as in Example 1.

[0082] 2.2 Reverse transcription

[0083] Reverse transcription using Primescript 1 st Strand cDNA synthesis kit, the operation steps are as follows:

[0084] (1) Add the following reaction liquid to the microcentrifuge tube, as shown in Table 1:

[0085] Table 1 Reaction liquid

[0086] reagent dose RNA 2.0μg dNTPs 1.0μl Oligo(dT) 2.0μl RNase free dH 2 O

Add to 10.0 μl

[0087] (2) Incubate at 70°C for 5 minutes, then rapidly cool ...

Embodiment 3

[0109] Example 3 PNLIPRP3 gene overexpression

[0110] 1. Construction of PNLIPRP3 gene recombinant plasmid

[0111] (1) Amplify the coding sequence of the PNLIPRP3 gene;

[0112] (2) Design amplification primers;

[0113] (3) The amplified PNLIPRP3 gene was ligated into the expression vector pcDNA3.0 to construct the pcDNA3.0-PNLIPRP3 recombinant expression vector.

[0114] 2. Culture and transfection of tongue squamous cell carcinoma

[0115] 2.1 Cell Culture

[0116] Human tongue squamous cell carcinoma cell line HN4 was cultured in DMEM / F12 medium containing 10% FBS, 100 U / mL penicillin and 100 μg / mL streptomycin. All cells were placed in 5% CO 2 in a 37°C cell incubator.

[0117] 2.2 Cell transfection

[0118] (1) 24h before transfection, cells were divided into 2 × 10 5 The cells were inoculated into 6-well plates, and DMEM / F12 medium was added, and the cells adhered overnight. When the cells reached 80-90% confluence, the cells were transfected.

[0119] (2) Di...

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Abstract

The invention discloses a PNLIPRP3 gene which can be adopted as a molecular marker for tongue squamous carcinoma diagnosis and treatment. Experiments show that compared with normal tissue, the PNLIPRP3 gene is low in expression quantity in tongue squamous carcinoma tissue. The invention further discloses application of the PNLIPRP3 gene for preparing a drug for tongue squamous carcinoma treatment. The invention provides a novel tongue squamous carcinoma clinic treatment method, and further provides a novel drug target for tongue squamous carcinoma treatment.

Description

technical field [0001] The present invention relates to the field of tumor diagnosis, treatment, and prognosis prediction, and more particularly, to a method for tumor diagnosis and prognosis prediction by detecting abnormality of PNLIPRP3; and a tumor therapeutic agent for activating PNLIPRP3 gene or protein. Background technique [0002] Oral squamous cell carcinoma is the most common malignant tumor of head and neck, accounting for about 3% of systemic tumors and 90% of oral tumors, while tongue cancer ranks first in the incidence of oral squamous cell carcinoma. Although the incidence of oral cancer in developed countries has shown a slow downward trend in recent years, the overall incidence has been increasing. Most of the patients are men over 40 years old, and the most common sites are the tongue, cheeks, and gums. The occurrence of tongue squamous cell carcinoma is a complex process with multiple factors, steps and stages. Although the etiology and related mechanis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/156G01N33/57407G01N33/68
Inventor 马翠常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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